Induced Pluripotent Stem Cell and Chromosome Core (iPSCC)

Established in 2006, the University of Connecticut (UConn) Stem Cell Core has substantially contributed to the Connecticut stem cell initiative by providing a central source of technologies and materials for research on human pluripotent stem cells: embryonic stem cells (hESC) and induced pluripotent stem cells (hiPSC). The stem cell core is a collaborative effort. Genome Editing, The Cell Bank, and Pluripotent Stem Cell Reprogramming are currently located at the UConn Health campus in Farmington. The Genetics and Genomics Division, including the Chromosome Core, is located at UConn Storrs campus. Single Cell Genomics is based at the Jackson Laboratory for Genomic Medicine in Farmington. The UConn Stem Cell Core is funded by UConn and Connecticut State Stem Cell Research Program. With these crucial services, many Connecticut researchers have developed novel approaches to addressing fundamental biological questions and tackling deleterious diseases.

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Contacts

Staff

Faculty Scientific Advisers

Location

Campus Address

Cell and Genome Sciences Building
UConn Health

Mailing Address

263 Farmington Avenue, MC 6022
Farmington CT 06032-6022

Services & Rates

Reprogramming Services

We provide integration-free reprogramming services using Sendai virus or episomal delivery method. Tissue samples accepted include skin fibroblast, peripheral blood, cord blood, PBMCs, and cells cultured from urine. Reprogramming can be done on feeder containing conditions or feeder independent conditions.

Specifications

Validation Services

Pluripotency Immunochemistry Test:

Immunostaining is included in standard iPSC derivation service. However, this service may also be ordered separately from our iPSC derivation service. Markers include Oct4 and Tra-1-60 or SSEA4
iPSCs are differentiated into embryoid bodies and specification of endoderm, ectoderm, and mesoderm lineages are assayed by qRT-PCR using the Applied Biosystems® TaqMan® hPSC Scorecard.
We provide a biochemical mycoplasma test using Myco-Alert kit®. Submit 1 - 2 ml of spent culture medium to be tested.

Specifications

Additional Services and Products

Biobanking Services:

We will perform QC measures, expand, store, and distribute your custom hESC and iPSC lines when requested by your collaborators or as a back-up storage for your lab.

Established hESC (H1 and H9, CT1 and CT2) and iPSC lines provided as frozen vials or live cultures
Validated Serum Replacement (KOSR) to make iPSC/hESC culture medium
bFGF aliquots, 50 mcg

We provide training in iPSC/hESC Basic Culture Techniques training as well as troubleshooting support.
- Workshops are available for iPSC/hESC differentiation to neural, cardiomyocyte, and vascular endothelial lineages.

Specifications

Karyotyping

Karyotype analysis of G-banded metaphase chromosomes will detect both numerical and structural chromosome abnormalities. Analysis of human metaphase chromosomes is done using standard protocols for chromosome harvesting, slide-making and G-banding in order to characterize cell lines for chromosome number and rearrangements at a 5-10Mb resolution.

Specifications

Cells should be submitted for karyotyping one day before the culture would normally be passaged to provide for a sufficient mitotic index (number of dividing cells). For each cell line submitted, a minimum of twenty-cells will be counted and sexed, five cells completely analyzed. A karyogram will be produced for two cells per clonal line. Additional counts will be performed in cases of suspected mosaicism. High-resolution banding at a 3-5Mb resolution, and special stains (C, Cd, NOR, and DA/DAPI) will be performed as indicated or requested.

Internal Rates

External Rates

Fluorescence In Situ Hybridization (FISH)

Fluorescence In Situ Hybridization (FISH) will detect the presence/absence or location of a specific gene or chromosome at ~ 100kb resolution. FISH studies are useful to demonstrate micro deletions or duplications and to demonstrate the presence of gene rearrangements. FISH also permits rapid detection of monosomies, trisomies, and numerical sex chromosome abnormalities. FISH involves the hybridization of a target DNA sequence labeled with a fluorescent dye (called a probe).

Specifications

Probes are currently available from a number of vendors or can be produced in our laboratory. Probes can be specific to unique sequences (e.g., ELN gene for Williams Syndrome on chromosome 7), or repetitive DNA (e.g., alpha-satellite centromere sequences), or chromosome paints (e.g., sequences that comprise chromosome 7). FISH analysis can be performed using one or multiple probes, provided the fluorescent labels for each probe are different and the microscope has the appropriate filter to detect the label. FISH can be performed on harvested metaphase cells, interphase cells, or on direct cell preparations. A FISH analysis includes examination of 20 metaphase cells or 100 interphase cells. Color fluorescence photomicrographs of representative FISH findings are provided.

Internal Rates

Affymetrix CytoScan HD SNP and CNV Analysis

Internal Rates

External Rates

$325	Karyotyping
$600	Karyotyping and FISH

$498	Karyotyping
$918	Karyotyping and FISH

$400/FISH	Based on single probe hybridization. Additional $50 for each additional probe, up to three probes hybridization maximum.
$600/FISH and Karyotyping
$1500/FISH with BAC prep of probe required

$1269	1 Sample
$1822	2 Samples
$2373	3 Samples
$2932	4 Samples
$3636	5 Samples

$1942	1 Sample
$2788	2 Samples
$3631	3 Samples
$4486	4 Samples
$5563	5 Samples

Induced Pluripotent Stem Cell and Chromosome Core (iPSCC)

Induced Pluripotent Stem Cell and Chromosome Core (iPSCC)

Contacts

Staff

Christopher Stoddard

Human Genome Editing Services

Noelle Germain, Ph.D.

Lab Manager

Yaling Liu

Research Assistant

Faculty Scientific Advisers

Judith Brown, Ph.D.

Assistant Professor in Residence

Rachel O'Neil, Ph.D.

Professor

Gordon Carmichael, Ph.D.

Professor

Location

Campus Address

Mailing Address

Services & Rates

Reprogramming Services

Validation Services

Additional Services and Products

Karyotyping

Fluorescence In Situ Hybridization (FISH)

Affymetrix CytoScan HD SNP and CNV Analysis